Effect of serum albumin supplementation on in vitro capacitation and fertilization of caprine oocytes |
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| Institution: | 1. C.I.R.G., Makhdoom, PO Farah-281122, Mathura (UP), India;2. College of Veterinary Science and A.H., Kumarganj, Faizabad (UP), India;1. College of Materials Science and Engineering, Chongqing University, Chongqing 400030, China;2. Chongqing Academy of Metrology and Quality Inspection, Chongqing 401121, China;1. College of Veterinary Medicine, University of Tennessee, Knoxville, TN, USA;2. School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA, USA;3. Department of Animal Science, University of Tennessee, Knoxville, TN, USA;4. School of Mathematical and Natural Sciences, Berry College, Mount Berry, GA, USA;1. Institute of Geotechnics, Slovak Academy of Sciences, 040 01 Košice, Slovakia;2. Institute of Material Science of Seville (CSIC-US), 410 92 Seville, Spain;3. Institute of Electronics and Photonics, Slovak University of Technology and International Laser Centre, 81219 Bratislava, Slovakia;1. Center for Molecular Prevention and Environmental Medicine, Department of Preventive Medicine, Tokai University School of Medicine, Shimokasuya, Isehara, Kanagawa, Japan;2. Central Institute for Experimental Animals, Tonomachi, Kawasaki-ku, Kawasaki, Kanagawa, Japan;3. Department of Neurology, Tokai University School of Medicine, Shimokasuya, Isehara, Kanagawa, Japan;4. Department of Molecular Life Science, Division of Basic Medical Science and Molecular Medicine, Tokai University School of Medicine, Shimokasuya, Isehara, Kanagawa, Japan;1. Physiology and Climatology Division, Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India;2. Division of Livestock Economics, Statistics and Information Technology, Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India;1. School of Microelectronics Science and Solid-State Electronics, University of Electronic Science and Technology, Chengdu 610054, China;2. School of Materials Science & Engineering, Guilin University of Electronic Technology, Guilin 541004, China;3. Department of Chemical Engineering, Monash University, Clayton, Vic 3800, Australia |
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| Abstract: | The aim of the investigation was to study the effect of purity and the type of serum albumin on in vitro fertilization (IVF) and cleavage rate of in vitro matured goat oocytes. Ovaries were collected from the local abattoir and transported within 4 h to the laboratory in warm saline (37 °C) containing 100 IU penicillin-G and 100 μg streptomycin sulfate per ml. A total of 2509 cumulus oocyte complexes (COCs) were collected from 1313 ovaries. Oocytes were matured in TCM-199 medium containing FSH (5 μg/ml), LH (5 μg/ml) and estradiol-17β (1 μg/ml), supplemented with 20% fetal bovine serum at 38.5 °C and 5% CO2 in an incubator under humidified air for 27 h. After 27 h of in vitro maturation (IVM), oocytes were denuded, washed and randomly divided into 4 groups. Group 1 consisted of in vitro matured oocytes (n = 627) co-incubated with sperm in a 50 μl drop of TALP medium containing a 6 mg/ml crystalline bovine serum albumin (BSA) fraction V and 10 μg/ml heparin. Group 2 was comprised of in vitro matured oocytes (n = 470), co-incubated with sperm in a 50 μl drop of TALP medium containing 3 mg/ml crystalline BSA fraction V, 10% estrous goat serum and 10 μg/ml heparin. Group 3 was comprised of in vitro matured oocytes (n = 489) co-incubated with sperm in a 50 μl drop of TALP medium containing a 6 mg/ml fatty acid free BSA and 10 μg/ml heparin. Group 4 consisted of in vitro matured oocytes (n = 422) co-incubated with sperm in a 50 μl drop of TALP medium containing 20% estrous goat serum and 10 μg/ml heparin. After 18 h of co-incubation, the oocyte–sperm mixture was washed in the culture medium 15–20 times and cultured in 50 μl EDM. Cleavage of the in vitro fertilized oocytes were recorded 48 h post-insemination under an inverted phase contrast microscope. The average oocyte recovery rate/ovary and maturation rate was 1.91% and 80.03%, respectively. The cleavage rate in Group 1, Group 2, Group 3 and Group 4 was 1.59%, 8.93%, 11.86% and 35.30%, respectively. It could be concluded that the use of fatty acid free albumin resulted in a significantly higher (P < 0.05) cleavage rate, compared to unmodified albumin, and the supplementation of 20% estrous goat serum in the fertilization medium, significantly (P < 0.05) increased the cleavage rate of in vitro matured goat oocytes, compared to defatted albumin. |
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